| Summary: | The working isntruction was created for separation vitamins A, D and E by means of monolithic column engaged in system sequentional injectional analysis. Following conditions of separation were chosen after performance of optimalization: Analysis at monolithic column 25 mm long Chromolith Flash RP-18e column, 4,6 mm I.D. (Merck, Germany), with speed (10 µl/s) with applied mobile phase acetonitril metanol (1:1, v/v). Monolithic column was in SIA system and was engaged between moreway valve and flow Z cell. Spectrophotometric detection was performed at wave length for absorbing maximum for vitamins A - 324 nm, D - 265 nm, E - 285. Total SIA analysis time was 400 s (from that 230 s lasted separation process). From basic chromatographic validation parameters were evaluated the following symetric factor As, resolution Rs, amount theoretical trays and repeatability. Symetric factor for vitamin A - 1.309; vitamin D2 - 1.302; vitamin D3 - 1.286; vitamin E - 1.158. Peak resolution for vitamins A-D2 je 3.607; D2-D3 4.232; D-E 1.532. Amount theoretical stages for vitamin A 998, vitamin D2 1621, vitamin D3 2091 and vitamin E 2376. Repeatability was worked out from 6 sprays, and relative standard deviation of particular substances was calculated from the highest point of the graph peak. Repeatability was for...
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